[Mature teratoma of the ovary--a clinicopathologic study of 214 teratomas of the ovary].
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Biomedical subjects
Publications and source records attributed to J C Chang.
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DNP-specific, class II-restricted cloned T cells were shown to kill DNP-bearing A20.2J (A20-DNP) antigen-presenting cells. This killing was DNP-specific and was restricted by IA. Results from bystander cytotoxicity, cold-target inhibition, and protein and lymphokine inhibition experiments indicated that killing of A20-DNP targets was mediated by direct lysis. In addition to the direct lysis, antigen stimulation of the T cells also resulted in production of a soluble cytolytic factor which killed bystander L929 fibroblast cells. This killing was sensitive to inhibition of protein synthesis and lymphokine production but was not affected by the addition of cold A20-DNP target cells. Additional studies showed that other antigen-presenting cells, i.e., DNP-bearing P388D1 and splenic macrophages, were also lysed by the cloned T cells. These findings may indicate that lysis of target cells by nominal antigen-specific, class II-restricted T cells plays a role in immune regulation and/or immune protection.
We performed morphological studies on the epidermal peridermal granules (PG) of newborn mice using histological, fluorescent and electron microscopic methods. PG appeared during the 17th to 18th day of foetal life, and remained in the outermost epidermal layers until 5 days after birth. PG were evenly distributed over the entire cutaneous surface of the mouse, except the tail. PG were found to contain protein and corresponded to sites of transglutaminase activity, but they were devoid of lipids and nucleic acids. They could be differentiated clearly from keratohyalin granules. Together, these studies suggest that PG represent a unique epidermal protein product that serves as a marker of late foetal development.
The synergistic effects of tobacco smoking and alcohol consumption on the incidence of upper respiratory cancer may be linked to their common ability to produce acetaldehyde, an irritant and potential mutagen. Since alcohol consumption in most individuals results in very low concentrations of acetaldehyde in the blood, we determined whether bronchopulmonary cellular components are capable of oxidizing ethanol to acetaldehyde. We found that significant production of acetaldehyde occurred in vitro after incubation of human bronchopulmonary washings with 25 mM ethanol. Acetaldehyde production was increased in active smokers and related to microorganisms in the bronchopulmonary tract. It was abolished by preincubation of the washings with antibiotics and was reproduced in vitro with Streptococcus pneumoniae. Normal pulmonary cells in bronchopulmonary washings did not produce acetaldehyde from ethanol.
Cathepsin B-like activity was determined in alveolar macrophages (AM) and cell-free bronchoalveolar lavage fluid (BALF) obtained from volunteers who were current cigarette smokers and compared with that found in lifetime nonsmokers. Enzyme activity was determined with benzyloxycarbonyl-Leu-Leu-Arg-2-naphthylamide as the substrate. Specific activity of the enzyme was more than twice as high in AM from smokers than in cells from nonsmokers (35,600 +/- 2,250 versus 16,000 +/- 860; p less than 0.001) and about 10 times as high in BALF from smokers than in that from nonsmokers (3,060 +/- 380 versus 300 +/- 25; p less than 0.001). Because cathepsin B is capable of degrading structural lung proteins and inactivating alpha-1-proteinase inhibitor, and the elastinolytic activity of AM may be mediated through cysteine proteinases such as cathepsin B, the finding of high concentrations of an enzyme with cathepsin B-like activity in AM and BALF from smokers suggests the need to explore the role of the enzyme in structural lung damage associated with cigarette smoking.
Granulomatous inflammation was induced in the lungs of rats for assessment of the suitability of this animal species for long-term study of granuloma development and resolution and for comparison of the histologic changes with the cellular profile in bronchoalveolar lavage (BAL) samples. It was found that after a single intravenous injection of bacillus Calmette-Guérin (BCG) organisms suspended in saline-0.01% Triton, complete Freund's adjuvant (CFA), or BCG suspended in CFA (BCG + CFA), distinct pathologic patterns developed in the lungs during the acute stages. BCG alone caused numerous small epithelioid granulomas associated with interstitial infiltration; CFA alone caused large discrete granulomas with minimal interstitial changes, and BCG + CFA caused large granulomas associated with diffuse interstitial infiltration. Following CFA or BCG + CFA the maximum number and size of granulomas were reached approximately 4 weeks after injection. By 8 weeks collagen was seen in the larger granulomas in sections stained with hematoxylin and eosin from animals given CFA or BCG + CFA. Maximum collagen deposition was seen at 16 weeks. From that point onward the degree of collagen deposition decreased, so that by approximately 42 weeks collagen was no longer seen. At 52 weeks residual granulomatous lesions consisted of a few foci of foamy macrophages surrounded by several layers of lymphocytes or epithelioid cells surrounded by dense layers of lymphocytes. Bronchoalveolar lavage samples revealed that at all time periods the number of leukocytes was increased and that the increase was due primarily to an influx of macrophages and lymphocytes. The increase in number of lymphocytes was so striking that at the peak of granulomatous changes following injection of BCG + CFA up to 50% of the total cells were found to be lymphocytes. The total number of leukocytes in BAL samples and the absolute number of lymphocytes closely paralleled the intensity of histologic changes seen in microscopic sections. It is concluded that the intravenous injection of BCG, CFA, or BCG + CFA in rats causes distinct and profound granulomatous patterns that are associated with increased cellularity in BAL samples. The findings suggest that the rat may be an excellent model for study of the mechanisms of granuloma development and resolution.
Protein amino groups can react with glucose without the aid of enzymes to form stable Amadori products containing 1-amino-1-deoxyketose residues. These adducts can undergo subsequent rearrangements and dehydrations to form various brown and fluorescent pigments. Recently, a chromophore, 2-(2-furoyl)-4(5)-(2-furanyl)-1H-imidazole (FFI), was isolated from acid hydrolysates of bovine serum albumin (BSA) and poly-L-lysine which had been incubated with glucose. To confirm the presence of FFI in situ, a radioimmunoassay was developed. A derivative of FFI, 4-furanyl-2-furoyl-1H-imidazole-1-hexanoic acid, was coupled to BSA and used to immunize rabbits. A radioactive FFI derivative was synthesized by reaction of 2-furyl-glyoxal with gamma-amino-[2,3-3H]butyric acid to form FFI-[3H]butyric acid. The resultant antiserum showed binding affinity to FFI and cross-reactivity for related compounds. FFI bound to proteins was liberated by acid hydrolysis or digestion by proteinase K prior to measurement. A linear relationship was seen between the amount of FFI equivalent detected and the amount of acid hydrolysate or enzymatic digest assayed. Poly-L-lysine and BSA incubated with glucose showed a time-dependent increase in the amounts of fluorescence and FFI equivalence. The detection of a time-related increase in the amount of FFI or a closely related structure in enzymatically digested proteins implicates it as an in situ product on proteins which have undergone the Maillard reaction with glucose. Of physiological significance is that FFI could also be detected in human globin and serum albumin from normal individuals. Thus, proteins exposed to glucose in vitro and in vivo form FFI as an in situ glycosylation product.
Highly sensitive and specific synthetic substrates were used to quantitate cathepsin B and D activity in peritoneal macrophages in response to stimulation in vivo with mineral oil and thioglycollate. After intraperitoneal instillation of mineral oil the activity of cathepsin B increased significantly (to 15 300 units/mg protein versus 7 340 in saline controls), reaching values approaching those found in alveolar macrophages (18 400 units/mg protein). Significantly greater stimulation of enzyme activity was obtained after intraperitoneal instillation of thioglycollate (23 600 units/mg protein). Cathepsin D activity also increased significantly after both mineral oil and thioglycollate. However, the increase was moderate (from 806 to about 1 200 units/mg protein), remaining still more than six times lower than in alveolar macrophages. The data are the first to demonstrate that cathepsin B activity can be stimulated in vivo in peritoneal macrophages by instillation of agents that induce acute inflammation. They also point to a differential control of expression of cathepsin B and D activity in both peritoneal and alveolar macrophages in spite of the common lysosomal origin of the two enzymes.
Survival was measured as a function of the dose of germicidal UV light for the bacteria Escherichia coli, Salmonella typhi, Shigella sonnei, Streptococcus faecalis, Staphylococcus aureus, and Bacillus subtilis spores, the enteric viruses poliovirus type 1 and simian rotavirus SA11, the cysts of the protozoan Acanthamoeba castellanii, as well as for total coliforms and standard plate count microorganisms from secondary effluent. The doses of UV light necessary for a 99.9% inactivation of the cultured vegetative bacteria, total coliforms, and standard plate count microorganisms were comparable. However, the viruses, the bacterial spores, and the amoebic cysts required about 3 to 4 times, 9 times, and 15 times, respectively, the dose required for E. coli. These ratios covered a narrower relative dose range than that previously reported for chlorine disinfection of E. coli, viruses, spores, and cysts.
Twenty-one patients with neoplastic fever due to malignancy were treated with naproxen. A prompt and complete lysis of fever was obtained in 20 patients within 12 hours when an adequate dose of naproxen was given, and a sustained normal temperature was maintained in all responding patients while receiving naproxen except for one in whom a low grade fever recurred. Lysis of fever usually was followed by excessive sweating and subjective symptomatic improvement. However, when naproxen was discontinued in ten patients, febrile state to the pretreatment level recurred in seven patients within three days. This observation suggests naproxen has a definite and effective antipyretic activity against neoplastic fever although it may recur as the drug is discontinued. Naproxen may be a useful adjunctive agent in patients with neoplastic fever for a short-term symptomatic relief.
Cloning and sequence analysis of the alpha-globin genes from a Sardinian patient with the nondeletion type of hemoglobin-H disease revealed a new type of thalassemia lesion. A mutation in the alpha 2-globin gene changes the initiation codon ATG to ACG and abolishes the function of this gene. Globin mRNA output from the affected alpha 2 locus is decreased relative to the alpha 1 locus. The mutation is detectable in genomic DNA by restriction analysis with the enzyme NcoI. Of the seven Sardinian patients with nondeletion alpha thalassemia screened with this enzyme, six had the initiation codon lesion.
Several new synthetic substrates fulfilling the specificity requirements of cathepsin D were synthesized. One of these D-Phe-Ser(O-CH2-C6H5)-Phe-Phe-Ala-Ala-pAB(pAB = p-aminobenzoate) proved to be highly sensitive and convenient for measuring activity. Enzyme determination was carried out in a two-step reaction. In the first step the enzyme hydrolyzes the Phe-Phe bond of the substrate at pH 3.4. In the second step aminopeptidase M (EC 3.4.11.2) degrades one of the products Phe-Ala-Ala-pAB at pH 7 to 8 with the release of free pAB, which is then determined by a diazotization procedure. Activity can be measured in as little as 1 to 5 micrograms of macrophage protein. The activity of cathepsin D in rat alveolar macrophages was almost ten times higher than in resident peritoneal macrophages, and more than 25 times higher than in blood monocytes. The data indicate that transformation of blood monocytes into macrophages is associated with a much greater increase of cathepsin D activity in alveolar than peritoneal macrophages.
The clinical utility of naproxen as an antipyretic agent was examined in the differential diagnosis of fever of undetermined origin in patients with cancer. Twenty-two patients with cancer and fever of undetermined origin for more than seven days were treated with naproxen to control fever when there was no evidence of infection after a careful initial evaluation, and in most cases, after failure of antibiotic therapy. In final analysis, none of five patients with infectious fever had responses to naproxen. In contrast, 14 of 15 patients with neoplastic fever showed a prompt, complete, and sustained lysis of fever within 24 hours after the initiation of naproxen treatment, and the patients also showed symptomatic improvement. One patient with neoplastic fever who did not have a response to naproxen had lysis of fever after the removal of necrotic tumor tissue. Two patients with fever from connective tissue disease had a partial lysis of fever in response to naproxen. These data suggest that naproxen specifically produces the lysis of neoplastic fever and, therefore, is a useful agent in assisting in the differential diagnosis of infectious fever and neoplastic fever in patients with cancer and fever of undetermined origin.
Inhalation of chlorine (Cl2) or formaldehyde (HCHO) stimulates the trigeminal nerve endings in the nasal mucosa and results in respiratory rate depression in a concentration-dependent manner. To determine tolerance and cross-tolerance, the concentration-response curves of respiratory depression were compared between naive rats and rats pre-exposed to Cl2 or HCHO. Chlorine tolerance development was time and concentration dependent, being significant following a 1-day (6 hr/day), 10 ppm exposure, and reaching the maximum in 4 days. At 2.5 ppm of Cl2, tolerance was significant only after 10 days of exposure. Rats tolerant to Cl2 also showed cross-tolerance to HCHO. Tolerance to HCHO was observed in rats exposed to 28 ppm for 4 days, but not in groups exposed to 15 ppm for 1, 4, or 10 days. However, significant cross-tolerance to Cl2 was evident following a 1-day exposure to 15 ppm HCHO, with greatest effect seen in the group exposed for 10 days. Tolerance was reduced after a 7-day recovery following a 4-day exposure. Cross-tolerance was reduced also, but to a much lesser extent. These results suggest a common mechanism for tolerance and cross-tolerance development, but different reactive sites may exist for Cl2 and HCHO at the trigeminal nerve endings.
Severe bleeding diathesis was the presenting problem in a patient with systemic amyloidosis characterized by generalized amyloid adenopathy. Epsilon amino caproic acid effectively reversed the fibrinolytic state with improvement of bleeding tendency and reduction of lymphadenopathy. The possible relationship and mechanisms of fibrinolysis are discussed.
In view of the differences that have been found between the most-probable-number and membrane filtration methods for the recovery of coliforms from chlorinated samples, the survival of total and fecal coliforms in UV-irradiated effluent samples, as tested by the most-probable-number and standard single-step membrane filtration methods, was compared. There were no significant differences in the survival of total and fecal coliforms, as tested by the two methods. In a separate set of experiments comparing total and fecal coliform survival, as tested by the most-probable-number method, only a very small but statistically significant difference of 0.1 log survival units was found. For UV-disinfected wastewater effluents, standard one-step membrane filtration procedures are comparable to standard most-probable-number procedures.
Although it has been demonstrated in several animal species that neutrophils aggregate in the pulmonary microvasculature after intravenous infusion of chemotactic factors or substances that activate the systemic complement system, studies in rabbits have revealed that affected neutrophils do not migrate out of capillaries into air spaces unless the infusion processes are combined with manipulative procedures involving the airways, such as intubation or instillation of an anesthetic agent. In this study, we report that after intravenous infusion of endotoxin (Escherichia coli) into Sprague-Dawley rats, in the absence of airway manipulation, the absolute number of neutrophils in bronchoalveolar lavage (BAL) samples increased significantly 24 and 48 h after injection. This represented nearly a 24-h delay from the time that maximal numbers of neutrophils were seen in lung tissue and approximately an 18-h delay from the time that maximal numbers appeared in peripheral blood. We also found that after infusion of endotoxin the number of pulmonary alveolar macrophages (PAM) recoverable in BAL samples fell dramatically within 2 h and remained suppressed for at least 12 h. We conclude that: (1) neutrophils are capable of migration into air spaces after aggregation in pulmonary capillaries in the absence of airway manipulative procedures or instillation of chemoattractants into air spaces and (2) endotoxemia affected surface parameters of PAM in vivo that determine recoverability by lavage.
Digestion of DNA from a patient with homozygous beta zero thalassemia from Calabria, Italy with the restriction endonuclease Mst II produced a pattern similar to the one obtained with sickle cell trait DNA in that the Mst II site at the beta 6 position on one chromosome was abolished. We cloned the DNA from this beta-thalassemia chromosome and performed sequence analysis. The deletion of a single nucleotide (A) at the GAG codon of the beta 6 position results in a frame shift and early beta-globin chain termination. This mutation occurs on a chromosome with a haplotype similar to two other Mediterranean beta-thalassemia lesions. The Mst II enzyme is useful for prenatal diagnosis of beta thalassemia in this population.